Search results for: genetic drift

Authors: Mercedes Bertotto, Marcelo Bello, Hector Goicoechea, Veronica Fusca

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The National Service of Agri-Food Health and Quality (SENASA), controls honey to detect contamination by synthetic or natural chemical substances and establishes and controls the traceability of the product. The utility of near-infrared spectroscopy for the detection of adulteration of honey with high fructose corn syrup (HFCS) was investigated. First of all, a mixture of different authentic artisanal Argentinian honey was prepared to cover as much heterogeneity as possible. Then, mixtures were prepared by adding different concentrations of high fructose corn syrup (HFCS) to samples of the honey pool. 237 samples were used, 108 of them were authentic honey and 129 samples corresponded to honey adulterated with HFCS between 1 and 10%. They were stored unrefrigerated from time of production until scanning and were not filtered after receipt in the laboratory. Immediately prior to spectral collection, honey was incubated at 40°C overnight to dissolve any crystalline material, manually stirred to achieve homogeneity and adjusted to a standard solids content (70° Brix) with distilled water. Adulterant solutions were also adjusted to 70° Brix. Samples were measured by NIR spectroscopy in the range of 650 to 7000 cm⁻¹. The technique of specular reflectance was used, with a lens aperture range of 150 mm. Pretreatment of the spectra was performed by Standard Normal Variate (SNV). The ant colony optimization genetic algorithm sample selection (ACOGASS) graphical interface was used, using MATLAB version 5.3, to select the variables with the greatest discriminating power. The data set was divided into a validation set and a calibration set, using the Kennard-Stone (KS) algorithm. A combined method of Potential Functions (PF) was chosen together with Partial Least Square Linear Discriminant Analysis (PLS-DA). Different estimators of the predictive capacity of the model were compared, which were obtained using a decreasing number of groups, which implies more demanding validation conditions. The optimal number of latent variables was selected as the number associated with the minimum error and the smallest number of unassigned samples. Once the optimal number of latent variables was defined, we proceeded to apply the model to the training samples. With the calibrated model for the training samples, we proceeded to study the validation samples. The calibrated model that combines the potential function methods and PLSDA can be considered reliable and stable since its performance in future samples is expected to be comparable to that achieved for the training samples. By use of Potential Functions (PF) and Partial Least Square Linear Discriminant Analysis (PLS-DA) classification, authentic honey and honey adulterated with HFCS could be identified with a correct classification rate of 97.9%. The results showed that NIR in combination with the PT and PLS-DS methods can be a simple, fast and low-cost technique for the detection of HFCS in honey with high sensitivity and power of discrimination.

Keywords: adulteration, multivariate analysis, potential functions, regression

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Authors: Indira K., Valsamma Joseph, I. S. Bright

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Introduction :Biofuels could replace fossil fuels and reduce our carbon footprint on the planet by technological advancements needed for sustainable and economic fuel production. Micro algae have proven to be a promising source to meet the current energy demand because of high lipid content and production of high biomass rapidly. Marine diatoms, which are key contributors in the biofuel sector and also play a significant role in primary productivity and ecology with high biodiversity and genetic and chemical diversity, are less well understood than other microalgae for producing hydrocarbons. Method :The marine diatom samples selected for hydrocarbon analysis were a total of eleven, out of which 9 samples were from the culture collection of NCAAH, and the remaining two of them were isolated by serial dilution method to get a pure culture from a mixed culture of microalgae obtained from the various cruise stations (350&357) FORV Sagar Sampada along the west coast of India. These diatoms were mass cultured in F/2 media, and the biomass harvested. The crude extract was obtained from the biomass by homogenising with n-hexane, and the hydrocarbons was further obtained by passing the crude extract through 500mg Bonna Agela SPE column and the quantitative analysis was done by GCHRMS analysis using HP-5 column and Helium gas was used as a carrier gas(1ml/min). The injector port temperature was 2400C, the detector temperature was 2500C, and the oven was initially kept at 600C for 1 minute and increased to 2200C at the rate of 60C per minute, and the analysis of a mixture of long chain hydrocarbons was done .Results:In the qualitative analysis done, the most potent hydrocarbon was found to be Psammodictyon Panduriforme (NCAAH-9) with a hydrocarbon mass of 37.27mg/g of the biomass and 2.1% of the total biomass 0f 1.395g and the other potent producer is Biddulphia(NCAAH 6) with hydrocarbon mass of 25.4mg/g of biomass and percentage of hydrocarbon is 1.03%. In the quantitative analysis by GCHRMS, the long chain hydrocarbons found in most of the marine diatoms were undecane, hexadecane, octadecane 3ethyl 5,2 ethyl butyl, Eicosane7hexyl, hexacosane, heptacosane, heneicosane, octadecane 3 methyl, triacontane. The exact mass of the long chain hydrocarbons in all the marine diatom samples was found to be Nonadecane 12C191H40, Tritriacontane,13-decyl-13-heptyl 12C501H102, Octadecane,3ethyl-5-(2-ethylbutyl 12C261H54, tetratetracontane 12C441H89, Eicosane, 7-hexyl 12C261H54. Conclusion:All the marine diatoms screened produced long chain hydrocarbons which can be used as diesel fuel with good cetane value example, hexadecane, undecane. All the long chain hydrocarbons can further undergo catalytic cracking to produce short chain alkanes which can give good octane values and can be used as gasoline. Optimisation of hydrocarbon production with the most potent marine diatom yielded long chain hydrocarbons of good fuel quality.

Keywords: biofuel, hydrocarbons, marine diatoms, screening

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Authors: Madison Hall

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The Assisted Colonization of Polar Bears to Antarctica requires a careful analysis of treaties to understand existing legal barriers to Ursus maritimus transport and movement. An absence of land-based migration routes prevent polar bears from accessing southern polar regions on their own. This lack of access is compounded by current treaties which limit human intervention and assistance to ford these physical and legal barriers. In a time of massive planetary extinctions, Assisted Colonization posits that certain endangered species may be prime candidates for relocation to hospitable environments to which they have never previously had access. By analyzing existing treaties, this paper will examine how polar bears are limited in movement by humankind’s legal barriers. International treaties may be considered codified reflections of anthropocentric values of the best knowledge and understanding of an identified problem at a set point in time, as understood through the human lens. Even as human social values and scientific insights evolve, so too must treaties evolve which specify legal frameworks and structures impacting keystone species and related biomes. Due to costs and other myriad difficulties, only a very select number of species will be given this opportunity. While some species move into new regions and are then deemed invasive, Assisted Colonization considers that some assistance may be mandated due to the nature of humankind’s role in climate change. This moral question and ethical imperative against the backdrop of escalating climate impacts, drives the question forward; what is the potential for successfully relocating a select handful of charismatic and ecologically important life forms? Is it possible to reimagine a different, but balanced Antarctic ecosystem? Listed as a threatened species under the U.S. Endangered Species Act, a result of the ongoing loss of critical habitat by melting sea ice, polar bears have limited options for long term survival in the wild. Our current regime for safeguarding animals facing extinction frequently utilizes zoos and their breeding programs, to keep alive the genetic diversity of the species until some future time when reintroduction, somewhere, may be attempted. By exploring the potential for polar bears to be relocated to Antarctica, we must analyze the complex ethical, legal, political, financial, and biological realms, which are the backdrop to framing all questions in this arena. Can we do it? Should we do it? By utilizing an environmental ethics perspective, we propose that the Ecological Commons of the Arctic and Antarctic should not be viewed solely through the lens of human resource management needs. From this perspective, polar bears do not need our permission, they need our assistance. Antarctica therefore represents a second, if imperfect chance, to buy time for polar bears, in a world where polar regimes, not yet fully understood, are themselves quickly changing as a result of climate change.

Keywords: polar bear, climate change, environmental ethics, Arctic, Antarctica, assisted colonization, treaty

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Authors: Rabab Makki

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Bone mineral density and bone metabolism are affected by various factors such as genetic, endocrine, mechanical and nutritional. Our understanding of nutritional influences on bone health is limited because most studies have focused on calcium. This study investigated the dietary factors which are likely t contribute to Osteoporosis in Saudi post-menopausal women, and correlated it with BMD. This is a case controlled study involved 36 postmenopausal Saudi females selected from the Orthopedics and osteoporosis outpatient clinics, and 25 postmenopausal Saudi females as controls from the primary clinic of Military Hospital in Riyadh. The women were diagnosed as osteoporotic based on the BMD measurement at any site (left femur neck, right femur neck, left total hip or right total hip or spine). Both the controls and the Osteoporotics were over 50 years of age and BMI between 31-34 kg/m2 had 2nd degree obesity, and were not free from other problems such as diabetes, hypertension, etc. Subjects (osteoporotics and controls) were interviewed to called data on demographic characterstics, medical history, dietary intake anthropometry (height and weight) bone mineral density. Blood samples were collected from subjects (Osteoporotics and controls). Analysis of serum calcium, vitamin D, phosphate were done at the main laboratory at Military Hospital Riyadh, by the laboratory technician while BMD was determined at the department of Nuclear Medicine by an expert technician and results were interpreted by radiologist.Data on frequency of consumption of animal food (meat, eggs, poultry and fish) and diary foods (milk, yogurt, cheese) of osteoporotic was less than control. In spite of the low intake there was no association with BMD.In general, the vegetables and fruits were consumed less by the osteoporotics than control. The only fruit which had shown a significant positive correlation is banana with right and left hip BMD total probably due to high potassium and minerals content which likely to prevent bone resorption. Mataziz vegetables combination of wheat showed a significant positive correlation with the same site (total right and left hip). Both osteoporotics abd controls were consuming table sugar. (But the sweet intake showed a significant negative correlation with left neck femur BMD, suggesting sucrose increase urinary calcium loss. Both osteoporotic and controls were consuming Arabic coffee. A negative significant correlation between intake of Arabic coffee and BMD of right neck femur of osteoporosis patient was observed. It could be suggested that increased intake of fruits and vegetables, might promote bone density while high intake of coffee and sugars might affect bone density, no significant correlation was observed between BMD at any site and diary product. We can say the major risk factors are inadequate nutrition. Further studies are needed among Saudi population to confirm these results.

Keywords: osteoporosi, Saudia Arabia, Riyadh Armed Forces, postmenopausal women

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Authors: Katarzyna Drela, Miroslaw Wielgos, Mikolaj Wrobel, Barbara Lukomska

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Mesenchymal stem cells (MSCs) have a great potential in regenerative medicine. Since the initial number of isolated MSCs is limited, in vitro propagation is often required to reach sufficient numbers of cells for therapeutic applications. During long-term culture MSCs may undergo genetic or epigenetic alterations that subsequently increase the probability of spontaneous malignant transformation. Thus, factors that influence genomic stability of MSCs following long-term expansions need to be clarified before cultured MSCs are employed for clinical application. The aim of our study was to investigate the potential for spontaneous transformation of human neonatal cord blood (HUCB-MSCs) and adult bone marrow (BM-MSCs) derived MSCs. Materials and Methods: HUCB-MSCs and BM-MSCs were isolated by standard Ficoll gradient centrifugations method. Isolated cells were initially plated in high density 106 cells per cm2. After 48 h medium were changed and non-adherent cells were removed. The malignant transformation of MSCs in vitro was evaluated by morphological changes, proliferation rate, ability to enter cell senescence, the telomerase expression and chromosomal abnormality. Proliferation of MSCs was analyzed with WST-1 reduction method and population doubling time (PDT) was calculated at different culture stages. Then the expression pattern of genes characteristic for mesenchymal or epithelial cells, as well as transcriptions factors were examined by RT-PCR. Concomitantly, immunocytochemical analysis of gene-related proteins was employed. Results: Our studies showed that MSCs from all bone marrow isolations ultimately entered senescence and did not undergo spontaneous malignant transformation. However, HUCB-MSCs from one of the 15 donors displayed an increased proliferation rate, failed to enter senescence, and exhibited an altered cell morphology. In this sample we observed two different cell phenotypes: one mesenchymal-like exhibited spindle shaped morphology and express specific mesenchymal surface markers (CD73, CD90, CD105, CD166) with low proliferation rate, and the second one with round, densely package epithelial-like cells with significantly increased proliferation rate. The PDT of epithelial-like populations was around 1day and 100% of cells were positive for proliferation marker Ki-67. Moreover, HUCB-MSCs showed a positive expression of human telomerase reverse transcriptase (hTERT), cMYC and exhibit increased number of CFU during the long-term culture in vitro. Furthermore, karyotype analysis revealed chromosomal abnormalities including duplications. Conclusions: Our studies demonstrate that HUCB-MSCs are susceptible to spontaneous malignant transformation during long-term culture. Spontaneous malignant transformation process following in vitro culture has enormous effect on the biosafety issues of future cell-based therapies and regenerative medicine regimens.

Keywords: mesenchymal stem cells, spontaneous, transformation, long-term culture

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Authors: Andy Ho

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Breast cancer is a common cancer in Australia. Early diagnosis is crucial for improving patient outcomes, as later-stage detection correlates with poorer prognoses. While multiparametric MRI offers superior sensitivity in detecting invasive and high-grade breast cancers compared to conventional mammography, its extended scan duration and high costs limit widespread application. As a result, full protocol MRI screening is typically reserved for patients at elevated risk. Recent advancements in imaging technology have facilitated the development of Abbreviated MRI protocols, which dramatically reduce scan times (<10 minutes compared to >30 minutes for full protocol). The potential for Abbreviated MRI to offer a more time- and cost-efficient alternative has implications for improving patient accessibility, reducing appointment durations, and enhancing compliance—especially relevant for individuals requiring regular annual screening over several decades. The purpose of this study is to assess the diagnostic efficacy of Abbreviated MRI for breast cancer screening among high-risk patients at the Royal Prince Alfred Hospital (RPA). This study aims to determine the sensitivity, specificity, and inter-reader variability of Abbreviated MRI protocols when interpreted by subspecialty-trained Breast Radiologists. A systematic review of the RPA’s electronic Picture Archive and Communication System identified high-risk patients, defined by Australian ‘Medicare Benefits Schedule’ criteria, who underwent Breast MRI from 2021 to 2022. Eligible participants included asymptomatic patients under 50 years old referred by the High-Risk Clinic due to a high-risk genetic profile or relevant familial history. The MRIs were anonymized, randomized, and interpreted by four Breast Radiologists, each independently completing standardized proforma evaluations. Radiological findings were compared against histopathology as the gold standard or follow-up imaging if biopsies were unavailable. Statistical metrics, including sensitivity, specificity, and inter-reader variability, were assessed. The Fleiss-Kappa analysis demonstrated a fair inter-reader agreement (kappa = 0.25; 95% CI: 0.19–0.32; p < 0.0001). The sensitivity for detecting malignancies was 0.75, with a specificity of 0.84. These findings underline the potential of Abbreviated MRI as a reliable screening tool for malignancies with significant specificity, though reduced sensitivity highlights the importance of robust radiologist training and consistent evaluation standards. Abbreviated MRI protocols exhibit promise as a viable screening option for high-risk patients, combining reduced scan times and acceptable diagnostic accuracy. Further work to refine interpretation practices and optimize training is essential to maximize the protocol’s utility in routine clinical screening and facilitate broader accessibility.

Keywords: abbreviated, breast, cancer, MRI

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Authors: Narges Bahmanyar, Masoud Ghorbani

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Rabies is caused by several species of the genus Lyssavirus in the Rhabdoviridae family. The disease is deadly encephalitis transmitted from warm-blooded animals to humans, and domestic and wild carnivores play the most crucial role in its transmission. The prevalence of rabies in poor areas of developing salinities is constantly posed as a global threat to public health. According to the World Health Organization, approximately 60,000 people die yearly from rabies. Of these, 60% of deaths are related to the Middle East. Although rabies encephalitis is incurable to date, awareness of the disease and the use of vaccines is the best way to combat the disease. Although effective vaccines are available, there is a high cost involved in vaccine production and management to combat rabies. Increasing the prevalence and discovery of new strains of rabies virus requires the need for safe, effective, and as inexpensive vaccines as possible. One of the approaches considered to achieve the quality and quantity expressed through the manufacture of recombinant types of rabies vaccine. Currently, livestock rabies vaccines are used only in inactivated or live attenuated vaccines, the process of inactivation of which pays attention to considerations. The rabies virus contains a negatively polarized single-stranded RNA genome that encodes the five major structural genes (N, P, M, G, L) from '3 to '5 . Rabies virus glycoprotein G, the major antigen, can produce the virus-neutralizing antibody. N-antigen is another candidate for developing recombinant vaccines. However, because it is within the RNP complex of the virus, the possibility of genetic diversity based on different geographical locations is very high. Glycoprotein G is structurally and antigenically more protected than other genes. Protection at the level of its nucleotide sequence is about 90% and at the amino acid level is 96%. Recombinant vaccines, consisting of a pathogenic subunit, contain fragments of the protein or polysaccharide of the pathogen that have been carefully studied to determine which of these molecules elicits a stronger and more effective immune response. These vaccines minimize the risk of side effects by limiting the immune system's access to the pathogen. Such vaccines are relatively inexpensive, easy to produce, and more stable than vaccines containing viruses or whole bacteria. The problem with these vaccines is that the pathogenic subunits may elicit a weak immune response in the body or may be destroyed before they reach the immune cells, which requires nanoparticles to overcome. Suitable for use as an adjuvant. Among these, biodegradable nanoparticles with functional levels are good candidates as adjuvants for the vaccine. In this study, we intend to use beta-glucan nanoparticles as adjuvants. The surface glycoprotein of the rabies virus (G) is responsible for identifying and binding the virus to the target cell. This glycoprotein is the major protein in the structure of the virus and induces an antibody response in the host. In this study, we intend to use rabies virus surface glycoprotein conjugated with beta-glucan nanoparticles to produce vaccines.

Keywords: rabies, vaccines, beta glucan, nanoprticles, adjuvant, recombinant protein

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Authors: Mathula Thangarajh

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Duchenne muscular dystrophy (DMD) is a severe form of X-linked muscular dystrophy caused by mutations in the dystrophin gene resulting in progressive skeletal muscle weakness. Boys with DMD also have significant cognitive disabilities. The intelligence quotient of boys with DMD, compared to peers, is approximately one standard deviation below average. Detailed neuropsychological testing has demonstrated that boys with DMD have a global developmental impairment, with verbal memory and visuospatial skills most significantly affected. Furthermore, the total brain volume and gray matter volume are lower in children with DMD compared to age-matched controls. These results are suggestive of a significant structural and functional compromise to the developing brain as a result of absent dystrophin protein expression. There is also some genetic evidence to suggest that mutations in the 3’ end of the DMD gene are associated with more severe neurocognitive problems. Our working hypothesis is that (i) boys with DMD do not make gains in neurodevelopmental skills compared to typically developing children and (ii) women carriers of DMD mutations may have subclinical cognitive deficits. We also hypothesize that there may be an intergenerational vulnerability of cognition, with boys of DMD-carrier mothers being more affected cognitively than boys of non-DMD-carrier mothers. The objectives of this study are: 1. Assess the neurodevelopment in boys with DMD at 4-time points and perform baseline neuroradiological assessment, 2. Assess cognition in biological mothers of DMD participants at baseline, 3. Assess possible correlation between DMD mutation and cognitive measures. This study also explores functional brain abnormalities in people with DMD by exploring how regional and global connectivity of the brain underlies executive function deficits in DMD. Such research can contribute to a better holistic understanding of the cognition alterations due to DMD and could potentially allow clinicians to create better-tailored treatment plans for the DMD population. There are four study visits for each participant (baseline, 2-4 weeks, 1 year, 18 months). At each visit, the participant completes the NIH Toolbox Cognition Battery, a validated psychometric measure that is recommended by NIH Common Data Elements for use in DMD. Visits 1, 3, and 4 also involve the administration of the BRIEF-2, ABAS-3, PROMIS/NeuroQoL, PedsQL Neuromuscular module 3.0, Draw a Clock Test, and an optional fMRI scan with the N-back matching task. We expect to enroll 52 children with DMD, 52 mothers of children with DMD, and 30 healthy control boys. This study began in 2020 during the height of the COVID-19 pandemic. Due to this, there were subsequent delays in recruitment because of travel restrictions. However, we have persevered and continued to recruit new participants for the study. We partnered with the Muscular Dystrophy Association (MDA) and helped advertise the study to interested families. Since then, we have had families from across the country contact us about their interest in the study. We plan to continue to enroll a diverse population of DMD participants to contribute toward a better understanding of Duchenne Muscular Dystrophy.

Keywords: neurology, Duchenne muscular dystrophy, muscular dystrophy, cognition, neurodevelopment, x-linked disorder, DMD, DMD gene

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Authors: Hanan Badr, Fahad Manee, Rao Shashidhar, Omar Bayoumy

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Introduction: Hypertension is the major risk factor for cardiovascular diseases and stroke and a universe leading cause of disability-adjusted life years and mortality. Adopting an unhealthy lifestyle is thought to be associated with developing hypertension regardless of predisposing genetic factors. This study aimed to examine the association between recreational physical activity (RPA), and sedentary behaviors with a risk of hypertension among ministry employees, where there is no role for occupational physical activity (PA), and to scrutinize participants’ time spent in RPA and sedentary behaviors on the working and weekend days. Methods: A cross-sectional study was conducted among randomly selected 2562 employees working at ten randomly selected ministries in Kuwait. To have a representative sample, the proportional allocation technique was used to define the number of participants in each ministry. A self-administered questionnaire was used to collect data about participants' socio-demographic characteristics, health status, and their 24 hours’ time use during a regular working day and a weekend day. The time use covered a list of 20 different activities practiced by a person daily. The New Zealand Physical Activity Questionnaire-Short Form (NZPAQ-SF) was used to assess the level of RPA. The scale generates three categories according to the number of hours spent in RPA/week: relatively inactive, relatively active, and highly active. Gender-matched trained nurses performed anthropometric measurements (weight and height) and measuring blood pressure (two readings) using an automatic blood pressure monitor (95% accuracy level compared to a calibrated mercury sphygmomanometer). Results: Participants’ mean age was 35.3±8.4 years, with almost equal gender distribution. About 13% of the participants were smokers, and 75% were overweight. Almost 10% reported doctor-diagnosed hypertension. Among those who did not, the mean systolic blood pressure was 119.9±14.2 and the mean diastolic blood pressure was 80.9±7.3. Moreover, 73.9% of participants were relatively physically inactive and 18% were highly active. Mean systolic and diastolic blood pressure showed a significant inverse association with the level of RPA (means of blood pressure measures were: 123.3/82.8 among relatively inactive, 119.7/80.4 among relatively active, and 116.6/79.6 among highly active). Furthermore, RPA occupied 1.6% and 1.8% of working and weekend days, respectively, while sedentary behaviors (watching TV, using electronics for social media or entertaining, etc.) occupied 11.2% and 13.1%, respectively. Sedentary behaviors were significantly associated with high levels of systolic and diastolic blood pressure. Binary logistic regression revealed that physical inactivity (OR=3.13, 95% CI: 2.25-4.35) and sedentary behaviors (OR=2.25, CI: 1.45-3.17) were independent risk factors for high systolic and diastolic blood pressure after adjustment for other covariates. Conclusions: Physical inactivity and sedentary lifestyle were associated with a high risk of hypertension. Further research to examine the independent role of RPA in improving blood pressure levels and cultural and occupational barriers for practicing RPA are recommended. Policies should be enacted in promoting PA in the workplace that might help in decreasing the risk of hypertension among sedentary occupation workers.

Keywords: physical activity, sedentary behaviors, hypertension, workplace

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Authors: Andrés Quiroz, Emilio Hormazabal, Ana Mutis, Fernando Ortega, Manuel Chacón-Fuentes, Leonardo Parra

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Red clover (Trifolium pratense L.) is an important forage species in temperate regions of the world. The main limitation of this species worldwide is a lack of persistence related to the high mortality of plants due to a complex of biotic and abiotic factors, determining a life span of two or three seasons. Because of the importance of red clover in Chile, a red clover breeding program was started at INIA Carillanca Research Center in 1989, with the main objective of improving the survival of plants, forage yield, and persistence. The main selection criteria for selecting new varieties have been based on agronomical parameters and biotic factors. The main biotic factor associated with red clover mortality in Chile is Hylastinus obscurus (Coleoptera: Curculionidae). Both larval and adults feed on the roots, causing weakening and subsequent death of clover plants. Pesticides have not been successful for controlling infestations of this root borer. Therefore, alternative strategies for controlling this pest are a high priority for red clover producers. Currently, the role of semiochemical in the interaction between H. obscurus and red clover plants has been widely studied for our group. Specifically, from the red clover foliage has been identified limonene is eliciting repellency from the root borer. Limonene is generated in the plant from two independent biosynthetic pathways, the mevalonic acid, and deoxyxylulose pathway. Mevalonate pathway enzymes are localized in the cytosol, whereas the deoxyxylulose phosphate pathway enzymes are found in plastids. In summary, limonene can be determinated by enzymatic bioassay using GPP as substrate and by limonene synthase expression. Therefore, the main objective of this work was to study genetic variation of limonene in material provided by INIA´s Red Clover breeding program. Protein extraction was carried out homogenizing 250 mg of leave tissue and suspended in 6 mL of extraction buffer (PEG 1500, PVP-30, 20 mM MgCl2 and antioxidants) and stirred on ice for 20 min. After centrifugation, aliquots of 2.5 mL were desalted on PD-10 columns, resulting in a final volume of 3.5 mL. Protein determination was performed according to Bradford with BSA as a standard. Monoterpene synthase assays were performed with 50 µL of protein extracts transferred into gas-tight 2 mL crimp seal vials after addition of 4 µL MgCl₂ and 41 µL assay buffer. The assay was started by adding 5 µL of a GPP solution. The mixture was incubated for 30 min at 40 °C. Biosynthesized limonene was quantified in a GC equipped with a chiral column and using synthetic R and S-limonene standards. The enzymatic the production of R and S-limonene from different Superqueli-Carillanca genotypes is shown in this work. Preliminary results showed significant differences in limonene content among the genotypes analyzed. These results constitute an important base for selecting genotypes with a high content of this repellent monoterpene towards H. obscurus.

Keywords: head space, limonene enzymatic determination, red clover, Hylastinus obscurus

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Authors: Benrithung Murry

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This paper highlights the biosocial determinants of health-seeking behavior in tribal population groups of northeast India, focusing on maternal and child health. The northeastern region of India is a conglomeration of several ethnic groups, most of which are scheduled as tribal groups. A total of 750 ever-married women in reproductive ages (15-49 years) were interviewed from three tribal groups of Nagaland, India using pre-tested and modified maternal health schedule. Data pertaining to reproductive performance of the mothers and their children health status were collected from 12 villages of Dimapur district, Nagaland, India. The sample for study comprises 212 Angami women, 267 Ao women, and 271 Sumi women, all of which belonging to tribal populations of Northeast India. Sex ratios of 15-49 years in these three populations are 1018.18, 1086.69, and 1106.92, respectively. 90% of the populations in the study are nuclear families, with about 10% of households falling below the poverty line as per the cutoffs for India. Female literacy level in these population groups is higher than the national average of 65.46%; however, about 30% of all married women are not engaged in any sort of earnings. Total fertility rates of these populations are alarming (Total Fertility Rate ≥ 6) and far from replacement fertility level, while infant mortality rates are found to be much lower than the national average of 34 per 1000. The perception and practice of maternal health in this region is unimpressive despite the availability of medical amenities. Only 3 % of mothers in the study have reported 4 times antenatal checkups during last two pregnancies. Other mothers have reported 1 to 3 times of antenatal checkups, but about 25% of them never visited a doctor during the entire pregnancy period. About 15% of mothers never took tetanus injection, while 40% of mothers never took iron folic supplements during pregnancy. Almost half of all women and their husbands do not use birth control measures even for the spacing of children, which has an immense impact on prenatal mortality mainly due to deliberate abortions: the percentage of prenatal mortality among Angami, Ao and Sumi populations is 44.88, 31.88 and 54.98, respectively per 1000 live births. The steep decline in fertility levels in most countries is a consequence of the increasing use of modern methods of contraception. However, among users of birth control measures in these populations, it is seen that most couples use it only after they have the desired number of children, thus its use having no substantial influence in reducing fertility. It is also seen that the majority of the children were only partially vaccinated. With many child deliveries being done at home, many newborns are not administered with polio at birth. Two-third of all children do not have complete basic immunization against polio, diphtheria, tetanus, pertussis, bacillus, and hepatitis besides others. Certain adherence to traditional beliefs and customs apart from the socio-economic factors is believed to have been operating in these populations, which determines their health-seeking behavior. While a more in-depth study combining biological, socio-cultural, economic, and genetic factors is suggested, there is an urgent need for intervention in these populations to combat with the poor maternal and child health status.

Keywords: case study, health behavior, mother and child, northeast india

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Authors: Anugrah Tripathi, H. S. Ginwal, Charul Kainthola

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Bamboos are considered as valuable resources which have the potential of meeting current economic, environmental and social needs. Bamboo has played a key role in humankind and its livelihood since ancient time. Distributed in diverse areas across the globe, bamboo makes an important natural resource for hundreds of millions of people across the world. In some of the Asian countries and northeast part of India, bamboo is the basis of life on many horizons. India possesses the largest bamboo-bearing area across the world and a great extent of species richness, but this rich genetic resource and its diversity have dwindled in the natural forest due to forest fire, over exploitation, lack of proper management policies, and gregarious flowering behavior. Bamboos which are well known for their peculiar, extraordinary morphology, show a lot of variation in many scales. Among the various bamboo species, Dendrocalamus strictus is the most abundant bamboo resource in India, which is a deciduous, solid, and densely tufted bamboo. This species can thrive in wide gradients of geographical as well as climatic conditions. Due to this, it exhibits a significant amount of variation among the populations of different origins for numerous morphological features. Morphological parameters are the front-line criteria for the selection and improvement of any forestry species. Study on the diversity among eight important morphological characters of D. strictus was carried out, covering 16 populations from wide geographical locations of India following INBAR standards. Among studied 16 populations, three populations viz. DS06 (Gaya, Bihar), DS15 (Mirzapur, Uttar Pradesh), and DS16 (Bhogpur, Pinjore, Haryana) were found as superior populations with higher mean values for parametric characters (clump height, no. of culms/ clump, circumference of clump, internode diameter and internode length) and with the higher sum of ranks in non-parametric characters (straightness, disease, and pest incidence and branching pattern). All of these parameters showed an ample amount of variations among the studied populations and revealed a significant difference among the populations. Variation in morphological characters is very common in a species having wide distribution and is usually evident at various levels, viz., between and within the populations. They are of paramount importance for growth, biomass, and quick production gains. Present study also gives an idea for the selection of the population on the basis of these morphological parameters. From this study on morphological parameters and their variation, we may find an overview of best-performing populations for growth and biomass accumulation. Some of the studied parameters also provide ideas to standardize mechanisms of selecting and sustainable harvesting of the clumps by applying simpler silvicultural systems so that they can be properly managed in homestead gardens for the community utilization as well as by commercial growers to meet the requirement of industries and other stakeholders.

Keywords: Dendrocalamus strictus, homestead garden, gregarious flowering, stakeholders, INBAR

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Authors: Luisa Müting, Wisnu Harto Adiwijoyo

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Agroforestry systems transform the agricultural landscapes in the Sahel region of Africa, providing food and farming products consumed for subsistence or sold for income. In the incrementally dry climate of the Sahel region, the spreading of agroforestry practices is integral for policymaker efforts to counteract land degradation and provide soil restoration in the region. Several measures on agroforestry practices have been implemented in the region by governmental and non-governmental institutions in recent years. However, despite the efforts, past research shows that awareness of how policies and interventions are being consumed and perceived by the public remains low. Therefore, interpreting public policy dilemmas by analyzing the public perception regarding agroforestry concepts and practices is necessary. Public perceptions and discourses can be an essential driver or constraint for the adoption of agroforestry practices in the region. Thus, understanding the public discourse behavior of crucial stakeholders could assist policymakers in developing inclusive and contextual policies that are relevant to the context of agroforestry adoption in Sahel region. To answer how information about agroforestry spreads and is perceived by the public. As internet usage increased drastically over the past decade, reaching a share of 33 percent of the population being connected to the internet, this research is based on online conversation data. Social media data from Facebook are gathered daily between April 2021 and April 2022 in Djibouti, Senegal, Mali, and Nigeria based on their share of active internet users compared to other countries in the Sahel region. A systematic methodology was applied to the extracted social media using discourse network analysis (DNA). This study then clustered the data by the types of agroforestry practices, sentiments, and country. Additionally, this research extracted the text data from online news media during the same period to pinpoint events related to the topic of agroforestry. The preliminary result indicates that tree management, crops, and livestock integration, diversifying species and genetic resources, and focusing on interactions and productivity across the agricultural system; are the most notable keywords in agroforestry-related conversations within the four countries in the Sahel region. Additionally, approximately 84 percent of the discussions were still dominated by big actors, such as NGO or government actors. Furthermore, as a subject of communication within agroforestry discourse, the Great Green Wall initiative generates almost 60 percent positive sentiment within the captured social media data, effectively having a more significant outreach than general agroforestry topics. This study provides an understanding for scholars and policymakers with a springboard for further research or policy design on agroforestry in the four countries of the Sahel region with systematically uncaptured novel data from the internet.

Keywords: sahel, djibouti, senegal, mali, nigeria, social networks analysis, public discourse analysis, sentiment analysis, content analysis, social media, online news, agroforestry, land restoration

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Authors: Cristina Opris, Bogdan Cojocaru, Madalina Tudorache, Simona M. Coman, Vasile I. Parvulescu, Camelia Bala, Bahir Duraki, Jeroen A. Van Bokhoven

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The tremendous achievements in the development of the society concretized by more sophisticated materials and systems are merely based on non-renewable resources. Consequently, after more than two centuries of intensive development, among others, we are faced with the decrease of the fossil fuel reserves, an increased impact of the greenhouse gases on the environment, and economic effects caused by the fluctuations in oil and mineral resource prices. The use of biomass may solve part of these problems, and recent analyses demonstrated that from the perspective of the reduction of the emissions of carbon dioxide, its valorization may bring important advantages conditioned by the usage of genetic modified fast growing trees or wastes, as primary sources. In this context, the abundance and complex structure of lignin may offer various possibilities of exploitation. However, its transformation in fuels or chemicals supposes a complex chemistry involving the cleavage of C-O and C-C bonds and altering of the functional groups. Chemistry offered various solutions in this sense. However, despite the intense work, there are still many drawbacks limiting the industrial application. Thus, the proposed technologies considered mainly homogeneous catalysts meaning expensive noble metals based systems that are hard to be recovered at the end of the reaction. Also, the reactions were carried out in organic solvents that are not acceptable today from the environmental point of view. To avoid these problems, the concept of this work was to investigate the synthesis of superparamagnetic core shell catalysts for the fragmentation of lignin directly in the aqueous phase. The magnetic nanoparticles were covered with a nanoshell of an oxide (niobia) with a double role: to protect the magnetic nanoparticles and to generate a proper (acidic) catalytic function and, on this composite, cobalt nanoparticles were deposed in order to catalyze the C-C bond splitting. With this purpose, we developed a protocol to prepare multifunctional and magnetic separable nano-composite Co@Nb2O5@Fe3O4 catalysts. We have also established an analytic protocol for the identification and quantification of the fragments resulted from lignin depolymerization in both liquid and solid phase. The fragmentation of various lignins occurred on the prepared materials in high yields and with very good selectivity in the desired fragments. The optimization of the catalyst composition indicated a cobalt loading of 4wt% as optimal. Working at 180 oC and 10 atm H2 this catalyst allowed a conversion of lignin up to 60% leading to a mixture containing over 96% in C20-C28 and C29-C37 fragments that were then completely fragmented to C12-C16 in a second stage. The investigated catalysts were completely recyclable, and no leaching of the elements included in the composition was determined by inductively coupled plasma optical emission spectrometry (ICP-OES).

Keywords: superparamagnetic core-shell catalysts, environmental production of fuels, renewable lignin, recyclable catalysts

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Authors: Vikas Kumar, Kailash Chandra, Kaomud Tyagi

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The insect order Thysanoptera includes small insects commonly called thrips. As insect vectors, only thrips are capable of Tospoviruses transmission (genus Tospovirus, family Bunyaviridae) affecting various crops. Currently, fifteen species of subfamily Thripinae (Thripidae) have been reported as vectors for tospoviruses. Frankliniella schultzei, which is reported as act as a vector for at least five tospovirses, have been suspected to be a species complex with more than one species. It is one of the historical unresolved issues where, two species namely, F. schultzei Trybom and F. sulphurea Schmutz were erected from South Africa and Srilanaka respectively. These two species were considered to be valid until 1968 when sulphurea was treated as colour morph (pale form) and synonymised under schultzei (dark form) However, these two have been considered as valid species by some of the thrips workers. Parallel studies have indicated that brown form of schultzei is a vector for tospoviruses while yellow form is a non-vector. However, recent studies have shown that yellow populations have also been documented as vectors. In view of all these facts, it is highly important to have a clear understanding whether these colour forms represent true species or merely different populations with different vector carrying capacities and whether there is some hidden diversity in 'Frankliniella schultzei species complex'. In this study, we aim to study the 'Frankliniella schultzei species complex' with molecular spectacles with DNA data from India and Australia and Africa. A total of fifty-five specimens was collected from diverse locations in India and Australia. We generated molecular data using partial fragments of mitochondrial cytochrome c oxidase I gene (mtCOI) and 28S rRNA gene. For COI dataset, there were seventy-four sequences, out of which data on fifty-five was generated in the current study and others were retrieved from NCBI. All the four different tree construction methods: neighbor-joining, maximum parsimony, maximum likelihood and Bayesian analysis, yielded the same tree topology and produced five cryptic species with high genetic divergence. For, rDNA, there were forty-five sequences, out of which data on thirty-nine was generated in the current study and others were retrieved from NCBI. The four tree building methods yielded four cryptic species with high bootstrap support value/posterior probability. Here we could not retrieve one cryptic species from South Africa as we could not generate data on rDNA from South Africa and sequence for rDNA from African region were not available in the database. The results of multiple species delimitation methods (barcode index numbers, automatic barcode gap discovery, general mixed Yule-coalescent, and Poisson-tree-processes) also supported the phylogenetic data and produced 5 and 4 Molecular Operational Taxonomic Units (MOTUs) for mtCOI and 28S dataset respectively. These results of our study indicate the likelihood that F. sulphurea may be a valid species, however, more morphological and molecular data is required on specimens from type localities of these two species and comparison with type specimens.

Keywords: DNA barcoding, species complex, thrips, species delimitation

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Authors: Fadwa Al Bawardi

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In March 2021, the Saudi Arabian Council of Ministers issued a decision to form a committee called the "Higher Committee for Research, Development and Innovation," a committee linked to the Council of Economic and Development Affairs, chaired by the Chairman of the Council of Economic and Development Affairs, and concerned with the development of the research, development and innovation sector in the Kingdom. In order to talk about the dimensions of this wonderful step, let us first try to answer the following questions. Is there a difference between creativity and innovation..? What are the factors of creativity in the individual. Are they mental genetic factors or are they factors that an individual acquires through learning..? The methodology included surveys that have been conducted on more than 500 individuals, males and females, between the ages of 18 till 60. And the answer is. "Creativity" is the creation of a new idea, while "Innovation" is the development of an already existing idea in a new, successful way. They are two sides of the same coin, as the "creative idea" needs to be developed and transformed into an "innovation" in order to achieve either strategic achievements at the level of countries and institutions to enhance organizational intelligence, or achievements at the level of individuals. For example, the beginning of smart phones was just a creative idea from IBM in 1994, but the actual successful innovation for the manufacture, development and marketing of these phones was through Apple later. Nor does creativity have to be hereditary. There are three basic factors for creativity: The first factor is "the presence of a challenge or an obstacle" that the individual faces and seeks thinking to find solutions to overcome, even if thinking requires a long time. The second factor is the "environment surrounding" of the individual, which includes science, training, experience gained, the ability to use techniques, as well as the ability to assess whether the idea is feasible or otherwise. To achieve this factor, the individual must be aware of own skills, strengths, hobbies, and aspects in which one can be creative, and the individual must also be self-confident and courageous enough to suggest those new ideas. The third factor is "Experience and the Ability to Accept Risk and Lack of Initial Success," and then learn from mistakes and try again tirelessly. There are some tools and techniques that help the individual to reach creative and innovative ideas, such as: Mind Maps tool, through which the available information is drawn by writing a short word for each piece of information and arranging all other relevant information through clear lines, which helps in logical thinking and correct vision. There is also a tool called "Flow Charts", which are graphics that show the sequence of data and expected results according to an ordered scenario of events and workflow steps, giving clarity to the ideas, their sequence, and what is expected of them. There are also other great tools such as the Six Hats tool, a useful tool to be applied by a group of people for effective planning and detailed logical thinking, and the Snowball tool. And all of them are tools that greatly help in organizing and arranging mental thoughts, and making the right decisions. It is also easy to learn, apply and use all those tools and techniques to reach creative and innovative solutions. The detailed figures and results of the conducted surveys are available upon request, with charts showing the %s based on gender, age groups, and job categories.

Keywords: innovation, creativity, factors, tools

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Authors: Parisa Shirzadeh

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Drug delivery systems in which drugs are traditionally used, multi-stage and at specified intervals by patients, do not meet the needs of the world's up-to-date drug delivery. In today's world, we are dealing with a huge number of recombinant peptide and protean drugs and analogues of hormones in the body, most of which are made with genetic engineering techniques. Most of these drugs are used to treat critical diseases such as cancer. Due to the limitations of the traditional method, researchers sought to find ways to solve the problems of the traditional method to a large extent. Following these efforts, controlled drug release systems were introduced, which have many advantages. Using controlled release of the drug in the body, the concentration of the drug is kept at a certain level, and in a short time, it is done at a higher rate. Graphene is a natural material that is biodegradable, non-toxic, and natural compared to carbon nanotubes; its price is lower than carbon nanotubes and is cost-effective for industrialization. On the other hand, the presence of highly effective surfaces and wide surfaces of graphene plates makes it more effective to modify graphene than carbon nanotubes. Graphene oxide is often synthesized using concentrated oxidizers such as sulfuric acid, nitric acid, and potassium permanganate based on Hummer 1 method. In comparison with the initial graphene, the resulting graphene oxide is heavier and has carboxyl, hydroxyl, and epoxy groups. Therefore, graphene oxide is very hydrophilic and easily dissolves in water and creates a stable solution. On the other hand, because the hydroxyl, carboxyl, and epoxy groups created on the surface are highly reactive, they have the ability to work with other functional groups such as amines, esters, polymers, etc. Connect and bring new features to the surface of graphene. In fact, it can be concluded that the creation of hydroxyl groups, Carboxyl, and epoxy and in fact graphene oxidation is the first step and step in creating other functional groups on the surface of graphene. Chitosan is a natural polymer and does not cause toxicity in the body. Due to its chemical structure and having OH and NH groups, it is suitable for binding to graphene oxide and increasing its solubility in aqueous solutions. Graphene oxide (GO) has been modified by chitosan (CS) covalently, developed for control release of doxorubicin (DOX). In this study, GO is produced by the hummer method under acidic conditions. Then, it is chlorinated by oxalyl chloride to increase its reactivity against amine. After that, in the presence of chitosan, the amino reaction was performed to form amide transplantation, and the doxorubicin was connected to the carrier surface by π-π interaction in buffer phosphate. GO, GO-CS, and GO-CS-DOX characterized by FT-IR, RAMAN, TGA, and SEM. The ability to load and release is determined by UV-Visible spectroscopy. The loading result showed a high capacity of DOX absorption (99%) and pH dependence identified as a result of DOX release from GO-CS nanosheet at pH 5.3 and 7.4, which show a fast release rate in acidic conditions.

Keywords: graphene oxide, chitosan, nanosheet, controlled drug release, doxorubicin

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Authors: Abdul Rehman, Bo Liu

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Axial flow turbines are commonly designed with high loads that generate strong secondary flows and result in high secondary losses. These losses contribute to almost 30% to 50% of the total losses. Non-axisymmetric endwall profiling is one of the passive control technique to reduce the secondary flow loss. In this paper, the non-axisymmetric endwall profile construction and optimization for the stator endwalls are presented to improve the efficiency of a high pressure turbine. The commercial code NUMECA Fine/ Design3D coupled with Fine/Turbo was used for the numerical investigation, design of experiments and the optimization. All the flow simulations were conducted by using steady RANS and Spalart-Allmaras as a turbulence model. The non-axisymmetric endwalls of stator hub and shroud were created by using the perturbation law based on Bezier Curves. Each cut having multiple control points was supposed to be created along the virtual streamlines in the blade channel. For the design of experiments, each sample was arbitrarily generated based on values automatically chosen for the control points defined during parameterization. The Optimization was achieved by using two algorithms i.e. the stochastic algorithm and gradient-based algorithm. For the stochastic algorithm, a genetic algorithm based on the artificial neural network was used as an optimization method in order to achieve the global optimum. The evaluation of the successive design iterations was performed using artificial neural network prior to the flow solver. For the second case, the conjugate gradient algorithm with a three dimensional CFD flow solver was used to systematically vary a free-form parameterization of the endwall. This method is efficient and less time to consume as it requires derivative information of the objective function. The objective function was to maximize the isentropic efficiency of the turbine by keeping the mass flow rate as constant. The performance was quantified by using a multi-objective function. Other than these two classifications of the optimization methods, there were four optimizations cases i.e. the hub only, the shroud only, and the combination of hub and shroud. For the fourth case, the shroud endwall was optimized by using the optimized hub endwall geometry. The hub optimization resulted in an increase in the efficiency due to more homogenous inlet conditions for the rotor. The adverse pressure gradient was reduced but the total pressure loss in the vicinity of the hub was increased. The shroud optimization resulted in an increase in efficiency, total pressure loss and entropy were reduced. The combination of hub and shroud did not show overwhelming results which were achieved for the individual cases of the hub and the shroud. This may be caused by fact that there were too many control variables. The fourth case of optimization showed the best result because optimized hub was used as an initial geometry to optimize the shroud. The efficiency was increased more than the individual cases of optimization with a mass flow rate equal to the baseline design of the turbine. The results of artificial neural network and conjugate gradient method were compared.

Keywords: artificial neural network, axial turbine, conjugate gradient method, non-axisymmetric endwall, optimization

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Authors: Yichao Liu, Peter Fransson, Julian Heidecke, Jonas Wallin, Joacim Rockloev

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Dengue, a mosquito-borne viral disease, poses a significant public health challenge in endemic tropical or subtropical countries, including Sri Lanka. To reveal insights into the complexity of the dynamics of this disease and study the drivers, a comprehensive model capable of both robust forecasting and insightful inference of drivers while capturing the co-circulating of several virus strains is essential. However, existing studies mostly focus on only one aspect at a time and do not integrate and carry insights across the siloed approach. While mechanistic models are developed to capture immunity dynamics, they are often oversimplified and lack integration of all the diverse drivers of disease transmission. On the other hand, purely data-driven methods lack constraints imposed by immuno-epidemiological processes, making them prone to overfitting and inference bias. This research presents a hybrid model that combines machine learning techniques with mechanistic modelling to overcome the limitations of existing approaches. Leveraging eight years of newly reported dengue case data, along with socioeconomic factors, such as human mobility, weekly climate data from 2011 to 2018, genetic data detecting the introduction and presence of new strains, and estimates of seropositivity for different districts in Sri Lanka, we derive a data-driven vector (SEI) to human (SEIR) model across 16 regions in Sri Lanka at the weekly time scale. By conducting ablation studies, the lag effects allowing delays up to 12 weeks of time-varying climate factors were determined. The model demonstrates superior predictive performance over a pure machine learning approach when considering lead times of 5 and 10 weeks on data withheld from model fitting. It further reveals several interesting interpretable findings of drivers while adjusting for the dynamics and influences of immunity and introduction of a new strain. The study uncovers strong influences of socioeconomic variables: population density, mobility, household income and rural vs. urban population. The study reveals substantial sensitivity to the diurnal temperature range and precipitation, while mean temperature and humidity appear less important in the study location. Additionally, the model indicated sensitivity to vegetation index, both max and average. Predictions on testing data reveal high model accuracy. Overall, this study advances the knowledge of dengue transmission in Sri Lanka and demonstrates the importance of incorporating hybrid modelling techniques to use biologically informed model structures with flexible data-driven estimates of model parameters. The findings show the potential to both inference of drivers in situations of complex disease dynamics and robust forecasting models.

Keywords: compartmental model, climate, dengue, machine learning, social-economic

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Authors: Siddhant Sethi, Yasuharu Takashima, Shigetaka Nakamura, Kenzo Fujimoto

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Site-directed mutagenesis is a renowned technique to introduce specific mutations in the genome. To achieve site-directed mutagenesis, many chemical and enzymatic approaches have been reported in the past like disulphite induced genome editing, CRISPR-Cas9, TALEN etc. The chemical methods are invasive whereas the enzymatic approaches are time-consuming and expensive. Most of these techniques are unusable in the cellular application due to their toxicity and other limitations. Photo-chemical cytosine deamination, introduced in 2010, is one of the major technique for enzyme-free single-point mutation of cytosine to uracil in DNA and RNA, wherein, 3-cyanovinylcarbazole nucleoside (CNVK) containing oligodeoxyribonucleotide (ODN) having CNVK at -1 position to that of target cytosine is reversibly crosslinked to target DNA strand using 366 nm and then incubated at 90ºC to accommodate deamination. This technique is superior to enzymatic methods of site-directed mutagenesis but has a disadvantage that it requires the use of high temperature for the deamination step which restricts its applicability in the in vivo applications. This study has been focused on improving the technique by reducing the temperature required for deamination. Firstly, the photo-cross-linker, CNVK has been modified by replacing cyano group attached to vinyl group with methyl ester (OMeVK), amide (NH2VK), and carboxylic acid (OHVK) to observe the acceleration in the deamination of target cytosine cross-linked to vinylcarbazole derivative. Among the derivatives, OHVK has shown 2 times acceleration in deamination reaction as compared to CNVK, while the other two derivatives have shown deceleration towards deamination reaction. The trend of rate of deamination reaction follows the same order as that of hydrophilicity of the vinylcarbazole derivatives. OHVK being most hydrophilic has shown highest acceleration while OMeVK is least hydrophilic has proven to be least active for deamination. Secondly, in the related study, the counter-base of the target cytosine, guanine has been replaced by inosine, 2-aminopurine, nebularine, and 5-nitroindole having distinct hydrogen bonding patterns with target cytosine. Among the ODNs with these counter bases, ODN with inosine has shown 12 fold acceleration towards deamination of cytosine cross-linked to CNVK at physiological conditions as compared to guanosine. Whereas, when 2-aminopurine, nebularine, and 5-nitroindole were used, no deamination reaction took place. It can be concluded that inosine has potential to be used as the counter base of target cytosine for the CNVK mediated photo-cross-linking induced deamination of cytosine. The increase in rate of deamination reaction has been attributed to pattern and number of hydrogen bonding between the cytosine and counter base. One of the important factor is presence of hydrogen bond between exo-cyclic amino group of cytosine and the counter base. These results will be useful for development of more efficient technique for site-directed mutagenesis for C → U transformations in the DNA/RNA which might be used in the living system for treatment of various genetic disorders and genome engineering for making designer and non-native proteins.

Keywords: C to U transformation, DNA editing, genome engineering, ultra-fast photo-cross-linking

Procedia PDF Downloads 235

Authors: Blaga Alexandra Cristina, Kloetzer Lenuta, Bompa Amalia Stela, Galaction Anca Irina, Cascaval Dan

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Vitamin B9 is an important member of vitamins B group, being a growth factor, important for making genetic material as DNA and RNA, red blood cells, for building muscle tissues, especially during periods of infancy, adolescence and pregnancy. Its production by biosynthesis is based on the high metabolic potential of mutant Bacillus subtilis, due to a superior biodisponibility compared to that obtained by chemical pathways. Pertraction, defined as the extraction and transport through liquid membranes consists in the transfer of a solute between two aqueous phases of different pH-values, phases that are separated by a solvent layer of various sizes. The pertraction efficiency and selectivity could be significantly enhanced by adding a carrier in the liquid membrane, such as organophosphoric compounds, long chain amines or crown-ethers etc., the separation process being called facilitated pertraction. The aim of the work is to determine the impact of the presence of two extractants/carriers in the bulk liquid membrane, i.e. di(2-ethylhexyl) phosphoric acid (D2EHPA) and lauryltrialkylmetilamine (Amberlite LA2) on the transport kinetics of vitamin B9. The experiments have been carried out using two pertraction equipments for a free liquid membrane or bulk liquid membrane. One pertraction cell consists on a U-shaped glass pipe (used for the dichloromethane membrane) and the second one is an H-shaped glass pipe (used for h-heptane), having 45 mm inner diameter of the total volume of 450 mL, the volume of each compartment being of 150 mL. The aqueous solutions are independently mixed by means of double blade stirrers with 6 mm diameter and 3 mm height, having the rotation speed of 500 rpm. In order to reach high diffusional rates through the solvent layer, the organic phase has been mixed with a similar stirrer, at a similar rotation speed (500 rpm). The area of mass transfer surface, both for extraction and for reextraction, was of 1.59x10-³ m2. The study on facilitated pertraction with the mixture of two carriers, namely D2EHPA and Amberlite LA-2, dissolved in two solvents with different polarities: n-heptane and dichloromethane, indicated the possibility to obtain the synergic effect. The synergism has been analyzed by considering the vitamin initial and final mass flows, as well as the permeability factors through liquid membrane. The synergic effect has been observed at low D2EHPA concentrations and high Amberlite LA-2 concentrations, being more important for the low-polar solvent (n-heptane). The results suggest that the mechanism of synergic pertraction consists on the reaction between the organophosphoric carrier and vitamin B9 at the interface between the feed and membrane phases, while the aminic carrier enhances the hydrophobicity of this compound by solvation. However, the formation of this complex reduced the reextraction rate and, consequently, affects the synergism related to the final mass flows and permeability factor. For describing the influences of carriers concentrations on the synergistic coefficients, some equations have been proposed by taking into account the vitamin mass flows or permeability factors, with an average deviations between 4.85% and 10.73%.

Keywords: pertraction, synergism, vitamin B9, Amberlite LA-2, di(2-ethylhexyl) phosphoric acid

Procedia PDF Downloads 275

Authors: Z. Giagkazoglou, D. Loukovitis, C. Gubili, A. Imsiridou

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Along with the increase in human population, demand for seafood has increased. Despite the strict labeling regulations that exist for most marketed species in the European Union, seafood substitution remains a persistent global issue. Food fraud occurs when food products are traded in a false or misleading way. Mislabeling occurs when one species is substituted and traded under the name of another, and it can be intentional or unintentional. Crustaceans are one of the most regularly consumed seafood in Greece. Shrimps, prawns, lobsters, crayfish, and crabs are considered a delicacy and can be encountered in a variety of market presentations (fresh, frozen, pre-cooked, peeled, etc.). With most of the external traits removed, products as such are susceptible to species substitution. DNA barcoding has proven to be the most accurate method for the detection of fraudulent seafood products. To our best knowledge, the DNA barcoding methodology is used for the first time in Greece, in order to investigate the labeling practices for crustacean products available in the market. A total of 100 tissue samples were collected from various retailers and markets across four Greek cities. In an effort to cover the highest range of products possible, different market presentations were targeted (fresh, frozen and cooked). Genomic DNA was extracted using the DNeasy Blood & Tissue Kit, according to the manufacturer's instructions. The mitochondrial gene selected as the target region of the analysis was the cytochrome c oxidase subunit I (COI). PCR products were purified and sequenced using an ABI 3500 Genetic Analyzer. Sequences were manually checked and edited using BioEdit software and compared against the ones available in GenBank and BOLD databases. Statistical analyses were conducted in R and PAST software. For most samples, COI amplification was successful, and species-level identification was possible. The preliminary results estimate moderate mislabeling rates (25%) in the identified samples. Mislabeling was most commonly detected in fresh products, with 50% of the samples in this category labeled incorrectly. Overall, the mislabeling rates detected by our study probably relate to some degree of unintentional misidentification, and lack of knowledge surrounding the legal designations by both retailers and consumers. For some species of crustaceans (i.e. Squila mantis) the mislabeling appears to be also affected by the local labeling practices. Across Greece, S. mantis is sold in the market under two common names, but only one is recognized by the country's legislation, and therefore any mislabeling is probably not profit-motivated. However, the substitution of the speckled shrimp (Metapenaus monoceros) for the distinct, giant river prawn (Macrobranchium rosenbergii), is a clear example of deliberate fraudulent substitution, aiming for profit. To our best knowledge, no scientific study investigating substitution and mislabeling rates in crustaceans has been conducted in Greece. For a better understanding of Greece's seafood market, similar DNA barcoding studies in other regions with increased touristic importance (e.g., the Greek islands) should be conducted. Regardless, the expansion of the list of species-specific designations for crustaceans in the country is advised.

Keywords: COI gene, food fraud, labelling control, molecular identification

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Authors: Eugene Aninagyei-Bonsu

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Introduction: Agriculture has been a cornerstone of human civilization, ensuring food security and livelihoods for billions of people worldwide. In recent decades, rapid advancements in technology have revolutionized the agricultural sector, offering innovative solutions to enhance productivity, sustainability, and efficiency. This abstract summarizes key findings from a research study that explores the impacts of technology in modern agriculture and its implications for future food production systems. Methodologies: The research study employed a mixed-methods approach, combining quantitative data analysis with qualitative interviews and surveys to gain a comprehensive understanding of the role of technology in agriculture. Data was collected from various stakeholders, including farmers, agricultural technicians, and industry experts, to capture diverse perspectives on the adoption and utilization of agricultural technologies. The study also utilized case studies and literature reviews to contextualize the findings within the broader agricultural landscape. Major Findings: The research findings reveal that technology plays a pivotal role in transforming traditional farming practices and driving innovation in agriculture. Advanced technologies such as precision agriculture, drone technology, genetic engineering, and smart irrigation systems have significantly improved crop yields, reduced environmental impact, and optimized resource utilization. Farmers who have embraced these technologies have reported increased productivity, enhanced profitability, and improved resilience to environmental challenges. Furthermore, the study highlights the importance of accessible and affordable technology solutions for smallholder farmers in developing countries. Mobile applications, sensor technologies, and digital platforms have enabled small-scale farmers to access market information, weather forecasts, and agricultural best practices, empowering them to make informed decisions and improve their livelihoods. The research emphasizes the need for targeted policies and investments to bridge the digital divide and promote equitable technology adoption in agriculture. Conclusion: In conclusion, this research underscores the transformative potential of technology in agriculture and its critical role in advancing sustainable food production systems. The findings suggest that harnessing technology can address key challenges facing the agricultural sector, including climate change, resource scarcity, and food insecurity. By embracing innovation and leveraging technology, farmers can enhance their productivity, profitability, and resilience in a rapidly evolving global food system. Moving forward, policymakers, researchers, and industry stakeholders must collaborate to facilitate the adoption of appropriate technologies, support capacity building, and promote sustainable agricultural practices for a more resilient and food-secure future.

Keywords: technology development in modern agriculture, the influence of information technology access in agriculture, analyzing agricultural technology development, analyzing of the frontier technology of agriculture loT

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Authors: Ramanpreet, Gursatej Gandhi

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Background: Dependence on electromagnetic radiations involved in communication and information technologies has incredibly increased in the personal and professional world. Among the numerous radiations, sources are fixed site transmitters, mobile phone base stations, and power lines beside indoor devices like cordless phones, WiFi, Bluetooth, TV, radio, microwave ovens, etc. Rather there is the continuous emittance of radiofrequency radiations (RFR) even to those not using the devices from mobile phone base stations. The consistent and widespread usage of wireless devices has build-up electromagnetic fields everywhere. In fact, the radiofrequency electromagnetic field (RF-EMF) has insidiously become a part of the environment and like any contaminant may pose to be health-hazardous requiring assessment. Materials and Methods: In the present study, cytogenetic damage was assessed using the Buccal Micronucleus Cytome (BMCyt) assay as a function of radiation exposure after Institutional Ethics Committee clearance of the study and written voluntary informed consent from the participants. On a pre-designed questionnaire, general information lifestyle patterns (diet, physical activity, smoking, drinking, use of mobile phones, internet, Wi-Fi usage, etc.) genetic, reproductive (pedigrees) and medical histories were recorded. For this, 24 hour-personal exposimeter measurements (PEM) were recorded for unrelated 60 healthy adults (40 cases residing in the vicinity of mobile phone base stations since their installation and 20 controls residing in areas with no base stations). The personal exposimeter collects information from all the sources generating EMF (TETRA, GSM, UMTS, DECT, and WLAN) as total RF-EMF uplink and downlink. Findings: The cases (n=40; 23-90 years) and the controls (n=20; 19-65 years) matched for alcohol drinking, smoking habits, and mobile and cordless phone usage. The PEM in cases (149.28 ± 8.98 mV/m) revealed significantly higher (p=0.000) electric field strength compared to the recorded value (80.40 ± 0.30 mV/m) in controls. The GSM 900 uplink (p=0.000), GSM 1800 downlink (p=0.000),UMTS (both uplink; p=0.013 and downlink; p=0.001) and DECT (p=0.000) electric field strength were significantly elevated in the cases as compared to controls. The electric field strength in the cases was significantly from GSM1800 (52.26 ± 4.49mV/m) followed by GSM900 (45.69 ± 4.98mV/m), UMTS (25.03 ± 3.33mV/m), DECT (18.02 ± 2.14mV/m) and was least from WLAN (8.26 ± 2.35mV/m). The higher significantly (p=0.000) increased exposure to the cases was from GSM (97.96 ± 6.97mV/m) in comparison to UMTS, DECT, and WLAN. The frequencies of micronuclei (1.86X, p=0.007), nuclear buds (2.95X, p=0.002) and cell death parameter (condensed chromatin cells) were significantly (1.75X, p=0.007) elevated in cases compared to that in controls probably as a function of radiofrequency radiation exposure. Conclusion: In the absence of other exposure(s), any cytogenetic damage if unrepaired is a cause of concern as it can cause malignancy. Larger sample size with the clinical assessment will prove more insightful of such an effect.

Keywords: Buccal micronucleus cytome assay, cytogenetic damage, electric field strength, personal exposimeter

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Authors: Zhao Gao, Eran Edirisinghe

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The production of an accurate sketch of a suspect based on a verbal description obtained from a witness is an essential task for most criminal investigations. The criminal investigation system employs specifically trained professional artists to manually draw a facial image of the suspect according to the descriptions of an eyewitness for subsequent identification. Within the advancement of Deep Learning, Recurrent Neural Networks (RNN) have shown great promise in Natural Language Processing (NLP) tasks. Additionally, Generative Adversarial Networks (GAN) have also proven to be very effective in image generation. In this study, a trained GAN conditioned on textual features such as keywords automatically encoded from a verbal description of a human face using an RNN is used to generate photo-realistic facial images for criminal investigations. The intention of the proposed system is to map corresponding features into text generated from verbal descriptions. With this, it becomes possible to generate many reasonably accurate alternatives to which the witness can use to hopefully identify a suspect from. This reduces subjectivity in decision making both by the eyewitness and the artist while giving an opportunity for the witness to evaluate and reconsider decisions. Furthermore, the proposed approach benefits law enforcement agencies by reducing the time taken to physically draw each potential sketch, thus increasing response times and mitigating potentially malicious human intervention. With publically available 'CelebFaces Attributes Dataset' (CelebA) and additionally providing verbal description as training data, the proposed architecture is able to effectively produce facial structures from given text. Word Embeddings are learnt by applying the RNN architecture in order to perform semantic parsing, the output of which is fed into the GAN for synthesizing photo-realistic images. Rather than the grid search method, a metaheuristic search based on genetic algorithms is applied to evolve the network with the intent of achieving optimal hyperparameters in a fraction the time of a typical brute force approach. With the exception of the ‘CelebA’ training database, further novel test cases are supplied to the network for evaluation. Witness reports detailing criminals from Interpol or other law enforcement agencies are sampled on the network. Using the descriptions provided, samples are generated and compared with the ground truth images of a criminal in order to calculate the similarities. Two factors are used for performance evaluation: The Structural Similarity Index (SSIM) and the Peak Signal-to-Noise Ratio (PSNR). A high percentile output from this performance matrix should attribute to demonstrating the accuracy, in hope of proving that the proposed approach can be an effective tool for law enforcement agencies. The proposed approach to criminal facial image generation has potential to increase the ratio of criminal cases that can be ultimately resolved using eyewitness information gathering.

Keywords: RNN, GAN, NLP, facial composition, criminal investigation

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Authors: Esra Sengul, R. G. Aktas, M. E. Sitar, H. Isan

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Hepatocellular carcinoma (HCC) is a hepatocellular tumor commonly found on the surface of the chronic liver. HepG2 is the most commonly used cell type in HCC studies. The main proteins remaining in the blood serum after separation of plasma fibrinogen are albumin and globulin. The fact that the albumin showed hepatocellular damage and reflect the synthesis capacity of the liver was the main reason for our use. Alpha-Fetoprotein (AFP) is an albumin-like structural embryonic globulin found in the embryonic cortex, cord blood, and fetal liver. It has been used as a marker in the follow-up of tumor growth in various malign tumors and in the efficacy of surgical-medical treatments, so it is a good protein to look at with albumins. We have seen the morphological changes of dimethyl sulfoxide (DMSO) on HepG2 and decided to investigate its biochemical effects. We examined the effects of DMSO, which is used in cell cultures, on albumin, AFP and total protein at low doses. Material Method: Cell Culture: Medium was prepared in cell culture using Dulbecco's Modified Eagle Media (DMEM), Fetal Bovine Serum Dulbecco's (FBS), Phosphate Buffered Saline and trypsin maintained at -20 ° C. Fixation of Cells: HepG2 cells, which have been appropriately developed at the end of the first week, were fixed with acetone. We stored our cells in PBS at + 4 ° C until the fixation was completed. Area Calculation: The areas of the cells are calculated in the ImageJ (IJ). Microscope examination: The examination was performed with a Zeiss Inverted Microscope. Daytime photographs were taken at 40x, 100x 200x and 400x. Biochemical Tests: Protein (Total): Serum sample was analyzed by a spectrophotometric method in autoanalyzer. Albumin: Serum sample was analyzed by a spectrophotometric method in autoanalyzer. Alpha-fetoprotein: Serum sample was analyzed by ECLIA method. Results: When liver cancer cells were cultured in medium with 1% DMSO for 4 weeks, a significant difference was observed when compared with the control group. As a result, we have seen that DMSO can be used as an important agent in the treatment of liver cancer. Cell areas were reduced in the DMSO group compared to the control group and the confluency ratio increased. The ability to form spheroids was also significantly higher in the DMSO group. Alpha-fetoprotein was lower than the values of an ordinary liver cancer patient and the total protein amount increased to the reference range of the normal individual. Because the albumin sample was below the specimen value, the numerical results could not be obtained on biochemical examinations. We interpret all these results as making DMSO a caretaking aid. Since each one was not enough alone we used 3 parameters and the results were positive when we refer to the values of a normal healthy individual in parallel. We hope to extend the study further by adding new parameters and genetic analyzes, by increasing the number of samples, and by using DMSO as an adjunct agent in the treatment of liver cancer.

Keywords: hepatocellular carcinoma, HepG2, dimethyl sulfoxide, cell culture, ELISA

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Authors: Su Mon Saw, Joe Rothnagel

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Short open reading frames (sORFs) located in 5’UTR of mRNAs are known as uORFs. Characterization of uORF-encoded peptides (uPEPs) i.e., a subset of short open reading frame encoded peptides (sPEPs) and their translation regulation lead to understanding of causes of genetic disease, proteome complexity and development of treatments. Existence of uORFs within cellular proteome could be detected by LC-MS/MS. The ability of uORF to be translated into uPEP and achievement of uPEP identification will allow uPEP’s characterization, structures, functions, subcellular localization, evolutionary maintenance (conservation in human and other species) and abundance in cells. It is hypothesized that a subset of sORFs are translatable and that their encoded sPEPs are functional and are endogenously expressed contributing to the eukaryotic cellular proteome complexity. This project aimed to investigate whether sORFs encode functional peptides. Liquid chromatography-mass spectrometry (LC-MS) and bioinformatics were thus employed. Due to probable low abundance of sPEPs and small in sizes, the need for efficient peptide enrichment strategies for enriching small proteins and depleting the sub-proteome of large and abundant proteins is crucial for identifying sPEPs. Low molecular weight proteins were extracted using SDS-PAGE from Human Embryonic Kidney (HEK293) cells and Strong Cation Exchange Chromatography (SCX) from secreted HEK293 cells. Extracted proteins were digested by trypsin to peptides, which were detected by LC-MS/MS. The MS/MS data obtained was searched against Swiss-Prot using MASCOT version 2.4 to filter out known proteins, and all unmatched spectra were re-searched against human RefSeq database. ProteinPilot v5.0.1 was used to identify sPEPs by searching against human RefSeq, Vanderperre and Human Alternative Open Reading Frame (HaltORF) databases. Potential sPEPs were analyzed by bioinformatics. Since SDS PAGE electrophoresis could not separate proteins <20kDa, this could not identify sPEPs. All MASCOT-identified peptide fragments were parts of main open reading frame (mORF) by ORF Finder search and blastp search. No sPEP was detected and existence of sPEPs could not be identified in this study. 13 translated sORFs in HEK293 cells by mass spectrometry in previous studies were characterized by bioinformatics. Identified sPEPs from previous studies were <100 amino acids and <15 kDa. Bioinformatics results showed that sORFs are translated to sPEPs and contribute to proteome complexity. uPEP translated from uORF of SLC35A4 was strongly conserved in human and mouse while uPEP translated from uORF of MKKS was strongly conserved in human and Rhesus monkey. Cross-species conserved uORFs in association with protein translation strongly suggest evolutionary maintenance of coding sequence and indicate probable functional expression of peptides encoded within these uORFs. Translation of sORFs was confirmed by mass spectrometry and sPEPs were characterized with bioinformatics.

Keywords: bioinformatics, HEK293 cells, liquid chromatography-mass spectrometry, ProteinPilot, Strong Cation Exchange Chromatography, SDS-PAGE, sPEPs

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Authors: Davide Gotti, Milan Prodanovic, Sergio Pinilla, David Muñoz-Torrero

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Since its proposal, the Doyle-Fuller-Newman (DFN) lithium-ion battery model has gained popularity in the electrochemical field. In fact, this model provides the user with theoretical support for designing the lithium-ion battery parameters, such as the material particle or the diffusion coefficient adjustment direction. However, the model is mathematically complex as it is composed of several partial differential equations (PDEs) such as Fick’s law of diffusion, the MacInnes and Ohm’s equations, among other phenomena. Thus, to efficiently use the model in a time-domain simulation environment, the selection of the discretization technique is of a pivotal importance. There are several numerical methods available in the literature that can be used to carry out this task. In this study, a comparison between the explicit Euler, Crank-Nicolson, and Chebyshev discretization methods is proposed. These three methods are compared in terms of accuracy, stability, and computational times. Firstly, the explicit Euler discretization technique is analyzed. This method is straightforward to implement and is computationally fast. In this work, the accuracy of the method and its stability properties are shown for the electrolyte diffusion partial differential equation. Subsequently, the Crank-Nicolson method is considered. It represents a combination of the implicit and explicit Euler methods that has the advantage of being of the second order in time and is intrinsically stable, thus overcoming the disadvantages of the simpler Euler explicit method. As shown in the full paper, the Crank-Nicolson method provides accurate results when applied to the DFN model. Its stability does not depend on the integration time step, thus it is feasible for both short- and long-term tests. This last remark is particularly important as this discretization technique would allow the user to implement parameter estimation and optimization techniques such as system or genetic parameter identification methods using this model. Finally, the Chebyshev discretization technique is implemented in the DFN model. This discretization method features swift convergence properties and, as other spectral methods used to solve differential equations, achieves the same accuracy with a smaller number of discretization nodes. However, as shown in the literature, these methods are not suitable for handling sharp gradients, which are common during the first instants of the charge and discharge phases of the battery. The numerical results obtained and presented in this study aim to provide the guidelines on how to select the adequate discretization technique for the DFN model according to the type of application to be performed, highlighting the pros and cons of the three methods. Specifically, the non-eligibility of the simple Euler method for longterm tests will be presented. Afterwards, the Crank-Nicolson and the Chebyshev discretization methods will be compared in terms of accuracy and computational times under a wide range of battery operating scenarios. These include both long-term simulations for aging tests, and short- and mid-term battery charge/discharge cycles, typically relevant in battery applications like grid primary frequency and inertia control and electrical vehicle breaking and acceleration.

Keywords: Doyle-Fuller-Newman battery model, partial differential equations, discretization, numerical methods

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Authors: Arsyam Mawardi, Sony Suhandono, Azzania Fibriani, Fifi Fitriyah Masduki

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Malaria is an infectious disease caused by Plasmodium sp. This disease has a high prevalence in Indonesia, especially in Jayapura. The vaccine that is currently being developed has not been effective in overcoming malaria. This is due to the high polymorphism in the Plasmodium genome especially in areas that encode Plasmodium surface proteins. Merozoite Surface Protein 1 (MSP1) Plasmodium falciparum is a surface protein that plays a role in the invasion process in human erythrocytes through the interaction of Glycophorin A protein receptors and sialic acid in erythrocytes with Reticulocyte Binding Proteins (RBP) and Duffy Adhesion Protein (DAP) ligands in merozoites. MSP1 can be targeted to be a specific antigen and predicted epitope area which will be used for the development of diagnostic and malaria vaccine therapy. MSP1 consists of 17 blocks, each block is dimorphic, and has been marked as the K1 and MAD20 alleles. Exceptions only in block 2, because it has 3 alleles, among others K1, MAD20 and RO33. These polymorphisms cause allelic variations and implicate the severity of patients infected P. falciparum. In addition, polymorphism of MSP1 in Jayapura isolates has not been reported so it is interesting to be further identified and projected as a specific antigen. Therefore, in this study, we analyzed the allele polymorphism as well as detected the MSP1 epitope antigen candidate on block 2 P. falciparum. Clinical samples of selected malaria patients followed the consecutive sampling method, examining malaria parasites with blood preparations on glass objects observed through a microscope. Plasmodium DNA was isolated from the blood of malarial positive patients. The block 2 MSP1 gene was amplified using PCR method and cloned using the pGEM-T easy vector then transformed to TOP'10 E.coli. Positive colonies selection was performed with blue-white screening. The existence of target DNA was confirmed by PCR colonies and DNA sequencing methods. Furthermore, DNA sequence analysis was done through alignment and formation of a phylogenetic tree using MEGA 6 software and insilico analysis using IEDB software to predict epitope candidate for P. falciparum. A total of 15 patient samples have been isolated from Plasmodium DNA. PCR amplification results show the target gene size about ± 1049 bp. The results of MSP1 nucleotide alignment analysis reveal that block 2 MSP1 genes derived from the sample of malarial patients were distributed in four different allele family groups, K1 (7), MAD20 (1), RO33 (0) and MSP1_Jayapura (10) alleles. The most commonly appears of the detected allele is MSP1_Jayapura single allele. There was no significant association between sex variables, age, the density of parasitemia and alel variation (Mann Whitney, U > 0.05), while symptomatic signs have a significant difference as a trigger of detectable allele variation (U < 0.05). In this research, insilico study shows that there is a new epitope antigen candidate from the MSP1_Jayapura allele and it is predicted to be recognized by B cells with 17 amino acid lengths in the amino acid sequence 187 to 203.

Keywords: epitope candidate, insilico analysis, MSP1 P. falciparum, polymorphism

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Authors: Anthony F. Peressini

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The concept of race has recently come prominently back into discussion in the context of medicine and medical science, along with renewed effort to biologize racial concepts. This paper argues that this renewed effort to biologize race by way of medicine and population genetics fail on their own terms, and more importantly, that the philosophical project of biologizing race ought to be recognized for what it is—a retrograde racial project—and abandoned. There is clear agreement that standard racial categories and concepts cannot be grounded in the old way of racial naturalism, which understand race as a real, interest-independent biological/metaphysical category in which its members share “physical, moral, intellectual, and cultural characteristics.” But equally clear is the very real and pervasive presence of racial concepts in individual and collective consciousness and behavior, and so it remains a pressing area in which to seek deeper understanding. Recent philosophical work has endeavored to reconcile these two observations by developing a “thin” conception of race, grounded in scientific concepts but without the moral and metaphysical content. Such “thin,” science-based analyses take the “commonsense” or “folk” sense of race as it functions in contemporary society as the starting point for their philosophic-scientific projects to biologize racial concepts. A “philosophic-scientific analysis” is a special case of the cornerstone of analytic philosophy: a conceptual analysis. That is, a rendering of a concept into the more perspicuous concepts that constitute it. Thus a philosophic-scientific account of a concept is an attempt to work out an analysis of a concept that makes use of empirical science's insights to ground, legitimate and explicate the target concept in terms of clearer concepts informed by empirical results. The focus in this paper is on three recent philosophic-scientific cases for retaining “race” that all share this general analytic schema, but that make use of “medical necessity,” population genetics, and human genetic clustering, respectively. After arguing that each of these three approaches suffers from internal difficulties, the paper considers the general analytic schema employed by such biologizations of race. While such endeavors are inevitably prefaced with the disclaimer that the theory to follow is non-essentialist and non-racialist, the case will be made that such efforts are not neutral scientific or philosophical projects but rather are what sociologists call a racial project, that is, one of many competing efforts that conjoin a representation of what race means to specific efforts to determine social and institutional arrangements of power, resources, authority, etc. Accordingly, philosophic-scientific biologizations of race, since they begin from and condition their analyses on “folk” conceptions, cannot pretend to be “prior to” other disciplinary insights, nor to transcend the social-political dynamics involved in formulating theories of race. As a result, such traditional philosophical efforts can be seen to be disciplinarily parochial and to address only a caricature of a large and important human problem—and thereby further contributing to the unfortunate isolation of philosophical thinking about race from other disciplines.

Keywords: population genetics, ontology of race, race-based medicine, racial formation theory, racial projects, racism, social construction

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